Summary: | Background : Cholesterol oxidase (CHO) is an enzyme that catalyzes cholesterol oxidation and produces hydrogen peroxide (H2O2). This enzyme is produced by certain pathogenic and non-pathogenic microorganisms and is a commercially important enzyme in the world which has found wide application in various industries. The aim of the study was to isolate native bacteria producing CHO and their identification using microbial, biochemical and molecular methods.
Materials and Methods: A total of 187 samples were collected from wastewater of leather and soap factories, soil, dairy products, and stagnant water. After the samples were cultured, the bacteria were identified using the microbial and biochemical tests. Then, the methods of colorimetric and browning of culture medium were applied to select the colonies producing CHO.PCR 16s rRNA technique was used for final confirmation of obtained strains.
Results: Of the 187 collected samples, two microbial samples were isolated, both of which from the soil samples. The results of the biochemical tests showed that the two bacteria were Rhodococcus. Both methods of colorimetric and browning of culture medium confirmed the activity of CHO. Molecular identification of these strains were also confirmed by sequencing 16s rRNA gene.
Conclusion: In this study, native bacteria producing CHO were isolated. These two bacteria were classified as Rhodococcus using morphological, biochemical and molecular methods.
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